PROJECT 2: The emerging fields of genomics and proteomics have led to a better comprehension of the pathophysiology of cancer and the identification of novel signaling pathways. These pathways offer novel 'targets'(e.g. Akt, MEK, mTOR and Receptor Tyrosine Kinases) which has led to the development of 'lead molecules'designed to inhibit the signaling derived from these pathways. However, this poses a tremendous challenge for selecting and/or validating these targets and for broad profiling of lead molecules for candidate selection. Molecular imaging technologies have the potential to address these scientific and technological challenges. The overall goal of Project 2 is to develop strategies wherein activation or inhibition of key pathways in tumor formation as well as in the response of tumors to therapies can be non-invasive imaged. Since targeted therapies often lead to tumor cytostasis (Gl arrest), we will in Aim 1 develop and test a non-invasive reporter for proliferation (entry of cells to S-phase of the cell cycle from G1). This reporter will be used to investigate the efficacy of four targeted therapeutic agents (PTK 787, a receptor Tyrosine Kinase (PDGFR) inhibitor;Perifosine, an AKT inhibitor;CCI 779, an mTOR inhibitor and Cl 1040, a MEK inhibitor). In Aim 2 we will use a non-invasive reporter for apoptosis to test the hypothesis that while targeted therapies may not induce apoptosis as single agents, in combination with other targeted therapies or with traditional therapies induction of apoptosis will correlate with efficacy and enhanced tumor control. In Aim 3, we will develop a reporter for Akt function and use it to test the ability of PTK 787, Cl 779, Cl 1040 and Perifosine to inhibit Akt activity. Each of the three molecular imaging approaches will be validated using traditional "gold standard" measures of function of these pathways (e.g. Western blots, immunohistochemistry, kinase assays). We believe that studies proposed in Project 2, will result in the development of tools that will be invaluable in testing the efficacy of targeted therapeutic agents as well as in optimization of their dose, schedule and development of the most efficacious combination therapies. Pub. Health: Overall, this research effort will provide the rationale for initiation of clinical trials with combinations of molecularly targeted therapies for the treatment of malignant brain tumors. In addition, imaging biomarkers for early assessment of treatment response will be identified and validated which will lead to individualization of patient treatment. University of Michigan Ann Arbor, Michigan PHS 398(Rev. 09/04) Page 173 Form Page 2 Principal Investigator/Program Director (Last, First, Middle): ROSS, Brian D. KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator. List all other key personnel in alphabetical order, last name first. Name eRA Commons User Name Organization Role on Project Rehemtulla, Alnawaz Alnawaz University of Michigan Project Leader Luker, Gary. gluker University of Michigan Co-Investigator OTHER SIGNIFICANT CONTRIBUTORS Name Organization Role on Project Human Embryonic Stem Cells Kl No Q Yes If the proposed project Involves human embryonic stem cells, list below the registration number of the specific cell line(s) from thefollowing list: http://stemcells.nih.qov/registrv/index.asp. Usecontinuationpages asneeded. If a specific line cannotbe referenced at this time, include a statement that one from the Registrywill be used. Cell Line Disclosure Permission Statement. Applicable to SSIR/STTROnly. SeeSB1R/STTRinstructions. l~1 Yes l~l No PHS 398 (Rev. 09/04) Page 174 Form Page 2-continued Number the following pages consecutively throughout the application. Do not use suffixes such as 4a, 4b. Principal Investigator/Program Director (Last, first, middle): Ross, Brian D. THROUGH DETAILED BUDGET FOR INITIAL BUDGET PERIOD FR[unreadable]M DIRECT COSTS ONLY Rehemtulla/Project 2 1 2/1 /2006 11/30/2007 PERSONNEL (Applicant organization only) % DOLLAR AMOUNTREQUESTED(omit cents) TYPE EFFORT INST. ROLE ON APPT. ON BASE SALARY FRINGE PROJECT NAME (months) PROJ. SALARY REQUESTED BENEFITS TOTALS Project Leader Rehemtulla, Alnawaz 12 30% $171,922 $51,577 $15,473 $67,050 Co- Luker, Gary Investigator 12 10% $183,500 $18,350 $5,505 $23,855 Research Bhojani, Mahaveer Associate 12 100% $57,500 $57,500 $17,250 $74,750 Research Griffin, Laura (Yrs 2-5) Associate 12 $37,885 Research Hamilton, Christin Associate 12 100% $40,896 $40,896 $12,269 $53,165 SUBTOT&HI_iOt x^[unreadable] $168,323 $50,497 $218,820 | CONSULTANT COSTS EQUIPMENT (Itemize) SUPPLIES (Itemize by category) Cell Culture Supplies $6,000 Disposable Supplies $3,500 Molecular Biology Reagents $5,000 Luciferin and Coelatrazine $22,000 $36,500 TRAVEL Attendance to 1 -2 Scientific Meetings per Year $1,500 PATIENT CARE COSTS INPATIENT OUTPATIENT ALTERATIONS AND RENOVATIONS (Itemize bycategory) OTHER EXPENSES (Itemize bycategory) Animal Purchases $15,000 Animal Care (SSF inc.) $30,000 Publication Charges $1,000 Histopathology $500 $46,500 CONSORTIUM/CONTRACTUAL COSTS DIRECT COSTS SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PEF (Item 7a, FacePage) $ 303,320 1 CONSORTIUM/CONTRACTUAL COSTS FACILITIES AND ADMINISTRATION COSTS TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD $ 303,320 1 SBIR/STTR Only: FEE REQUESTED PHS 398 (Rev. 09/04) Page 175 Form Page 4